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Viña 2008 Apio
Viña 2008 Apio
Viña 2008 Apio
Original article
Effect of heat treatment and refrigerated storage on antioxidant
properties of pre-cut celery (Apium graveolens L.)
Summary This work studies the eect of two types of heat treatment, dry air at 48 C for 1 h and water immersion at
50 C for 90 s, and of storage time at 0 C on a number of quality parameters for pre-cut celery: browning
potential, soluble phenols content, total avonoids, chlorogenic acid, ascorbic acid and antioxidant capacity.
Pre-cut celery was placed in crystal polyethylene terephthalate trays covered with polyvinyl chloride lm.
Samples were taken after 0, 1, 7, 14 and 21 storage days. Treatments reduced browning potential and
chlorogenic acid content and, in addition, allowed ascorbic acid concentration to be retained for a longer
time. For this reason, the application of heat treatments in minimally processed celery would be benecial.
Keywords Antioxidant activity, Apium graveolens L., controlled stress, fresh cut vegetables, phenolics.
doi:10.1111/j.1365-2621.2006.01380.x
2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund
Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves 45
antioxidant activity and may help to protect cells against With regard to air heat treatments, they were carried
the oxidative damage caused by free radicals (Wada & out after applying chlorinated water to samples as
Ou, 2002). mentioned earlier. Once disinfested, the product was
Celery adapts easily to minimal processing but the treated in a heated air oven for combinations of 48 C
main detrimental factors for its quality are vascular 60 min and 50 C20 min, followed by storage at 0 C for
browning at the ends of cut petioles, aring of the cut 28 days. Once air treatments were nished, samples were
ends and development of pithiness (i.e. the formation of allowed to cool at room temperature before packaging.
aerenchyma in the pith) (Saltveit & Mangrich, 1996; To evaluate and select the diverse treatments, their
Loaiza-Velarde et al., 2003). It has been shown that a eect on sensorial attributes and damage development
heat-shock treatment can diminish wound-induced phy- was considered (specially rot, yellowing and softening).
siological changes leading to reduced quality (i.e. tissue
browning) and shortened shelf life (Loaiza-Velarde
Selected treatments and storage conditions
et al., 2003).
The objective of the present work was to analyse the The selected treatments to be studied in this work are: (i)
inuence of two types of heat treatment and of control sample (C), i.e. not exposed to heat treatment;
refrigerated storage on several chemical components (ii) thermally treated product by immersion in water at
contributing to the antioxidant power of pre-cut celery. 50 C for 90 s (I, immersion); (iii) thermally treated
product in dry-heated air (oven) at 48 C for 1 h (HA,
hot air).
Materials and methods
In all tests, trays 17-cm long, 13-cm wide and 5-cm
deep were used, which were made of crystal polyethylene
Plant material and processing
terephthalate (PET) covered with self-adhering polyvi-
Celery plants (Apium graveolens L.) cv Golden Boy, nyl chloride (PVC) lm (thickness, 10 lm; O2 permeab-
grown in greenhouse, were received from a La Plata ility, 11 232 cm3 m)2 atm)1 day)1; CO2 permeability,
grower (Province of Buenos Aires, Argentina). This is a 48 552 cm3 m)2 atm)1 day)1; water vapour permeability,
white or self-whitening variety, widely cultivated in the 40 g m)2 day)1). The trays contained 175 g of pro-
zone. Once the plants reached the commercial size (after duct and were kept for 3 weeks in a cold store at 0 C
about 2 months of being transplanted), they were with a relative humidity of 85%. Samples were taken
harvested early in the morning, brought to the laborat- for analysis at 0, 1, 7, 14, and 21 days. The whole ex-
ory and processed immediately. Leaves and basal periment was repeated twice.
segments of the rosettes were eliminated to obtain
unbranched petioles. They were washed in running
Chemical analysis
drinking water to remove any soil residues, and subse-
quently cut with a sharpened knife in 4-cm long sticks. For each sampling point, the material coming from
These were disinfected by immersion in chlorinated three trays was combined and homogenised. Immedi-
water (100 ppm active chlorine, pH 66.5, 8 C) for ately before the analysis, part of the pool was frozen in
3 min and blotted dry. liquid N2 and crushed in a laboratory mill (Janke &
Kunkel Ika Labortechnik A10, Staufen, Germany).
From this material, subsamples of exact weight were
Selection of treatments
taken to carry out the corresponding determinations.
For immersion heat treatments, the following prelim-
inary temperaturetime combinations were tested: Browning potential
45 C120 s, 50 C90 s and 55 C60 s, the storage Extraction was performed with ethanol 96 and absorb-
period being of 6 days at 20 C to speed up the ance (320 nm) of the solutions was measured (Loaiza-
manifestation of damage. Based on these results, a Velarde et al., 1997). Extractions and determinations
second stage of testing comprised immersion at 50 C were carried out in duplicate and nal results were
90 s and 55 C30 s, with a storage time at 0 C of expressed as absorbance units (AU) per gram of fresh
28 days. Immersion treatments were carried in heated tissue.
distilled water using a thermostatic bath with perma-
nent stirring. Celery cuts were placed in a plastic Total phenols content
basket, and dipped during the selected times. Samples Aliquots (20 mL) of the alcoholic extracts were concen-
were subsequently immersed in chlorinated water with trated at reduced pressure (30 mm Hg, 40 C) in a rotary
ice (100 ppm of active chlorine, pH 66.5) for 3 min evaporator R-124 (Buchi Labortechnik AG, Flawil,
for cooling and disinfestation. The product was pack- Switzerland), until dryness. Residues were resuspended
aged after eliminating the excess water by draining on in doubly distilled water. Total phenols were quantied
absorbent paper. employing the Folin-Ciocalteu reagent (Swain & Hillis,
2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund International Journal of Food Science and Technology 2008
46 Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves
1959), whereas absorbance readings were taken at column. In this determination, however, the mobile
760 nm. Catechin was used as standard in the 3.75 phase was a 70:30 mixture of acetonitrile:water with
12.75 lg mL)1 concentration range. Duplicate extrac- 0.01 m NH4H2PO4 and pH adjusted to 4.3 with ortho-
tions and determinations were conducted and nal phosphoric acid. Flow rate was 2 mL min)1, detection
results were expressed as lmol g)1 of fresh tissue. being carried out at 254 nm. For identication and
quantication, a standard ascorbic acid solution of
Chlorogenic acid concentration 35 lg mL)1 was employed. Extractions and determina-
This determination was carried out as reported in a tions were carried out in duplicate and nal results were
previous work (Vina & Chaves, 2006). Samples were expressed as milligram of ascorbic acid per 100 g of
extracted and concentrated as mentioned earlier. Here, fresh tissue.
residues were resuspended in 1 mL high-performance
liquid chromatograph (HPLC) grade methanol, and Antioxidant power
analysed in a Waters Model 6000A (Milford, MA, Samples previously frozen in N2 and crushed were
USA) HPLC, tted with UVVIS detector. A C18 treated with 5 mL of methanol. The antioxidant power
column was employed (particle diameter 5 lm; internal (AP) of the extracts was determined by reaction with the
diameter 4.6 mm; length 25 cm), using an 85:10:5 stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) in
mixture of water:methanol:formic acid as running sol- a methanol solution, using a modied version of the
vent. A ow rate of 1 mL min)1 was used. Detection method proposed by Brand-Williams et al. (1995).
was conducted at 320 nm. A standard solution of Concentration of the extracts was varied in the reaction
chlorogenic acid with a concentration of 0.87 lg mL)1 mixtures adding 0, 200, 400, 600, 800 or 1000 lL of each
was used both to identify and quantify this compound. of them to a 3.9 mL methanol solution of DPPH
The UVVIS spectrum of the fraction resulting from (25 ppm), completing a nal volume of 4.9 mL with
chromatographic runs was compared with the standard methanol. The reaction was allowed to progress and
solution to conrm identication. Extractions and absorbance was measured at 515 nm after a constant
determinations were conducted in duplicate and results value was reached. Then, DPPH was calculated
were expressed as nmol g)1 of fresh tissue. through a calibration straight line obtained in a range
of concentrations of this substance. Finally, the remain-
Total avonoids content ing DPPH concentration was plotted as a function
It was determined by the technique described by Kim of the extract volume in the reaction mixture, to
et al. (2003), with modications. Samples were extrac- calculate EC50 (eective mean concentration) for
ted, concentrated and resuspended in doubly distilled each sampling point. EC50 was dened as the mass
water as described earlier. To prepare reaction mixtures, (grams) of tissue required to reduce DPPH concentra-
a test tube was added with 1500 lL of doubly distilled tion to half its initial value. Extractions and determina-
water and 500 lL of the concentrated samples. Other tions were carried out in duplicate. Final results were
compounds were added sequentially: initially (zero time) expressed as AP, dened as the reciprocal of EC50
a volume of 150 lL of 5% NaNO2; after 5 min, 150 lL (AP 1/EC50).
of 10% AlCl3 and nally, after additional 6 min, 500 lL
of 1 m NaOH. Solutions were mixed by stirring in a
Statistical analysis
vortex and then absorbance at 510 nm was measured. A
standard curve was constructed based on catechin All data were treated by analysis of variance (anova).
concentrations in the range of 7.536.6 lg mL)1. Sources of variation were time (ve levels) and treat-
Extractions and determinations were conducted in ment (three levels). Means were compared using Fishers
duplicate. Total avonoid levels in the samples were least signicant dierence (LSD) test. Dierences at
expressed as nmol g)1 of fresh tissue. P < 0.05 were considered signicant.
International Journal of Food Science and Technology 2008 2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund
Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves 47
Table 1 Incidence of damage in pre-cut celery treated by water Initial values averaged 0.08 AU per gram of fresh tissue.
immersion or hot air after 28 storage days at 0 C A signicant increase in browning potential was
observed for control samples over the rst week of
Temperature and Soft rot
Treatment time combination incidence (%) Yellowing Softening
storage (P < 0.05), reaching a maximum at day 7, of
almost twice the initial value. From then on, there was a
Control 3 ++ tendency for browning to decrease, though nevertheless
Immersion 50 C90 s 4 the browning potential in the untreated product stored
55 C30 s 8 + at 0 C for 21 days was signicantly higher (P < 0.05)
Hot air 48 C1 h 4 + than its initial level. In immersion-treated samples,
50 C20 min 7 ++ +
browning potential was mostly constant (P > 0.05)
, unaffected product; +, moderately affected product; ++, seriously over the rst two weeks in cold store, to reduce towards
affected product. Percentage of soft rot incidence was calculated by the day 21, to values slightly below the initial value. Samples
number of pieces affected related to the total number of inspected pieces treated in hot air experienced a signicant increase
in each sampling point. (P < 0.05) of browning potential up to 14 days of
storage. At this time, a maximum of 1.4 times of the
50 C for 90 s was chosen; besides the exposure time of initial value was reached.
the 55 C treatment was reduced from 60 to 30 s. Again, In our experiments, the initial browning potential
the treatment with the best results was 50 C90 s values were 2.5 times as low as those found by Loaiza-
(Table 1). With respect to heated air treatments, the Velarde et al. (2003). These authors published browning
48 C1 h combination caused lower yellowing and potentials of 0.2 AU per gram of fresh tissue for petiole
minimised rmness losses (Table 1). pieces 5 mm in length, increasing up to 0.6 AU per gram
For these reasons, the treatments selected to study fresh tissue after 5 weeks at 0 C. Besides, for immer-
their eect on the antioxidant properties of pre-cut sion-treated samples at 50 C for 90 s, the increase in
celery were immersion in water at 50 C for 90 s and browning potential was delayed by 3 weeks compared
heated air at 48 C for 1 h. with the controls (Loaiza-Velarde et al., 2003), showing
a similar trend as that observed in our experiments,
where the immersion treatment hindered browning. Our
Browning potential results showed, in turn, that the hot air treatment was
Figure 1 shows the results in this topic. No signicant less eective. In spite of the preceding facts, enzymatic
dierences of browning were observed between the browning in the variety under study here (Golden Boy)
controls and the thermally treated pieces immediately did not produce severe damage because, although there
after heat stress by immersion or dry air application. was some development in the stored product (control
and treated), manifestations were highly localised
(brown-orange spots coincident with exposed vascular
0.16 strands).
2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund International Journal of Food Science and Technology 2008
48 Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves
International Journal of Food Science and Technology 2008 2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund
Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves 49
40.0
Hot air Time (days) C I HA
35.0
0 3.6 3.1 3.4
30.0 7 4.9 3.6 4.2
14 3.0 3.0 3.5
25.0 21 2.9 3.1 3.3
20.0 C, control; I, immersion thermal treatment (50 C, 90 s); HA, hot air
thermal treatment (48 C, 1 h).
15.0
2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund International Journal of Food Science and Technology 2008
50 Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves
International Journal of Food Science and Technology 2008 2007 The Authors. Journal compilation 2007 Institute of Food Science and Technology Trust Fund
Antioxidants in heat-treated pre-cut celery S. Z. Vina and A. R. Chaves 51
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